微环境在体外扩增内皮祖细胞过程中的作用
利用脐静脉内皮细胞或晚期内皮祖细胞作为滋养层细胞能够大幅度扩增晚期内皮祖细胞，并能形成血管状结构。摄影显微镜观察到在扩增单一干细胞时滋养层细胞参与细胞的不对称分裂。
延伸：对内皮祖细胞及其生长微环境的研究，有助于了解血管损伤修复过程。内皮祖细胞在心脑血管疾病、外周血管疾病、肿瘤血管形成及创伤愈合等方面均发挥重要作用，并为缺血性疾病的研究治疗提供了新思路。而最新的一项研究证明血管疾病的真凶是干细胞的异化（http://www.nature.com/ncomms/journal/v3/n6/full/ncomms1867.html）。对内皮祖细胞扩增方法及其微环境的研究有助于对血管疾病的了解与治疗。
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2012 Jun;34(3):202-6. doi: 10.3881/j.issn.1000-503X.2012.03.002.
Role of microenvironment in the process of expansion of late endothelial progenitor cell in vitro.
Wu LH, Song ZX, Liu XH, Li SZ, Han ZC, Uzan G.
Source
Department of Internal Medicine, Institute of Hematology and Hospital of Blood Disease, CAMS and PUMC, Tianjin 300020, China. [email protected]
Abstract
OBJECTIVE:
To study the role of the feeder layer cells as niche in the process of expansion of late endothelial progenitor cell in vitro.
METHODS:
We cultured mononuclear cells (MNC)from human peripheral blood (PB)on the plate with the feeder layer cells which were irradiated late endothelial progenitor cells(EPC)or human umbilical vein endothelial cells (HUVEC) by EGM-2. After 21 days, the numbers of obtained late EPC colonies were counted separately, and their surface antigen of the late EPC was verified by fluorescence-activated cell sorter (FACS) analysis, and their ability of forming vessel structure with Matrigel in vitro. The differentiation of single stem cell on the feeder layer cell was traced by video-microscopy.
RESULTS:
After 21 days of culture,(40.0±3.9)and(39.3±3.1)late EPC colonies that MNC of a hundred milliliter PB were cultured, respectively, on the feeder layer cells of EPC and HUVEC were much more than (2.0±1.3) colonies cultured on without the feeder layer cells (all P <0.05). These cells also expressed CD31, CD34, eNOS, FLt-1, P1H12, Sendo,VEcadherin, and CD117, as shown by FACS analysis. Furthermore, they formed vessel structure with Matrigel in vitro. The video-microscopy showed the asymmetric cell division was participated by the feeder layer cell during the expansion of single stem cell.
CONCLUSION:
The massive expansion of late EPC can be achieved by the provision of the feeder layer cells, which may be involved in the stem cell asymmetric cell division.
